The goal of this project is to develop a new highly sensitive and cost effective RNA Enzymatic Luminescence Assay (qELA) for high-throughput detection and quantification of microRNA in biological samples. The assay implements the same detection concept known from pyrosequencing, yet expanding pyrosequencing detection methodology for highly sensitive and accurate quantification of small RNA molecules. The proposed assay can match the performance of the real time quantitative PCR technique (rt-qPCR) in many applications and is simpler and more cost effective. The technology can be implemented in a number of commercial assays for application in life science research, drug discovery, and clinical diagnosis. PUBLIC HEALTH RELEVANCE: The goal of this project is to develop a new high performance cost effective bioluminescence assay for microRNA analysis. The proposed assay provides technologically advanced and significantly less expensive alternative for real time polymerase chain reaction technology, which currently dominates in the market. The proposed bioluminescent assay is universal and can be used in various applications in life science research, drug discovery, clinical diagnosis by 1,800 clinical laboratories and thousands of life science research laboratories in US.